Time-domain/ELDOR EPR Spectrometer

时域/ELDOR EPR 光谱仪

• 批准号: 7596125
• 负责人: ALEX I. SMIRNOV
• 金额: $ 50万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596125
• 关键词: 20 year old; Applications Grants; Area; Binding; Biology; Catalytic Domain; Chemistry; Development; Drug Delivery Systems; Electronics; Enzymes; Funding; Hybrids; Ions; Label; Measures; Membrane Proteins; Metals; Methodology; Methods; North Carolina; Phospholipids; Physiologic pulse; Principal Investigator; Protein Microchips; Proteins; Public Health; RNA; Recovery; Relaxation; Research; Research Personnel; Research Project Grants; Structure; System; Time; Training; Training Programs; Transfer RNA; Universities; Virion; enzyme structure; graduate student; instrument; instrumentation; lipid transfer protein; nanoparticle; nanoscale; public health relevance; research study; self assembly

DESCRIPTION (provided by applicant): This is a Shared Instrumentation Grant proposal for the acquisition of a modern commercial CW and FT-EPR Bruker EleXsys X-band spectrometer for use by a core group of six established Principal Investigators and one beginning investigator from Universities in the Research Triangle area of North Carolina. Currently, there is no such instrument in the Triangle area. The ongoing research projects involve applications of modern time-domain EPR methods including pulsed saturation recovery and HYSCORE to a wide range of biomedical problems and systems from hybrid nanoscale assemblies for membrane protein biochips to lipid transfer proteins, self-assembly of tRNA and drug-delivery assisted by nanoparticle-templated virions, and the structures of the enzymes' catalytic sites. Currently, all these projects are either pursued with available continuous wave X-band EPR instruments that are at least 20 years old or in collaborating with other groups that have Bruker EleXsys X-band spectrometer available. With this proposed major upgrade all seven core research projects will benefit from recent developments in instrumentation and methodology of time-domain EPR methods at X-band. Specifically, we will be able to 1) obtain long-range (to up to 60-70 angstrom) distance constrains for large biomolecular assemblies and partially folded structures; 2) directly measure electronic relaxation rates of specifically labeled proteins, RNAs, and phospholipids in the presence and absence of paramagnetic relaxation agents in order to gain valuable structural information; and 3) obtain unique information on paramagnetic metal ion coordination upon substrate binding to catalytic enzymes (HYSCORE experiment).PUBLIC HEALTH RELEVANCE: The relevance of this project to public health will be in providing primarily NIH-funded investigators as well as other investigators from NCSU, UNC, and the Research Triangle area with an advanced FT- EPR spectrometer as these investigators develop new projects involving structure-function studies of proteins and RNAs. The new FT-EPR spectrometer will also enhance training of graduate students from the newly established NCSU RNA Biology/Chemistry -Biology Interface Training Program that involves ten NCSU Departments.

描述（由申请人提供）：这是一项共享仪器赠款提案，旨在购买一台现代商用 CW 和 FT-EPR Bruker EleXsys X 波段光谱仪，供由 6 名已建立的首席研究员和一名来自各大学的初级研究员组成的核心小组使用。北卡罗来纳州的研究三角地区。目前，三角地区还没有这样的仪器。正在进行的研究项目涉及现代时域 EPR 方法（包括脉冲饱和恢复和 HYSCORE）在广泛的生物医学问题和系统中的应用，从膜蛋白生物芯片的混合纳米级组装到脂质转移蛋白、tRNA 自组装和药物输送借助纳米颗粒模板病毒粒子和酶催化位点的结构。目前，所有这些项目要么使用已有至少 20 年历史的连续波 X 波段 EPR 仪器，要么与拥有 Bruker EleXsys X 波段光谱仪的其他团队合作。通过这项拟议的重大升级，所有七个核心研究项目都将受益于 X 波段时域 EPR 方法的仪器和方法的最新发展。具体来说，我们将能够 1) 获得大型生物分子组装体和部分折叠结构的长程（高达 60-70 埃）距离约束； 2）在存在和不存在顺磁弛豫剂的情况下直接测量特定标记的蛋白质、RNA和磷脂的电子弛豫率，以获得有价值的结构信息； 3) 获得关于底物与催化酶结合时顺磁金属离子配位的独特信息（HYSCORE 实验）。 公共健康相关性：该项目与公共健康的相关性主要在于提供 NIH 资助的研究人员以及 NCSU 的其他研究人员、北卡罗来纳大学和三角研究区配备了先进的 FT-EPR 光谱仪，这些研究人员正在开发涉及蛋白质和 RNA 结构功能研究的新项目。新的 FT-EPR 光谱仪还将加强对新成立的 NCSU RNA 生物学/化学-生物学界面培训项目（涉及 NCSU 十个系）的研究生的培训。

项目成果

Oxidation of Pyrrole by Dehaloperoxidase-Hemoglobin: Chemoenzymatic Synthesis of Pyrrolin-2-Ones.

脱卤过氧化物酶-血红蛋白对吡咯的氧化：吡咯啉-2-酮的化学酶合成。

• DOI: 10.1039/c7cy00781g
• 发表时间: 2017
• 期刊: Catalysis science & technology
• 影响因子: 5
• 作者: McCombs NL;Smirnova T;Ghiladi RA
• 通讯作者: Ghiladi RA

Peptide-Membrane Interactions by Spin-Labeling EPR.

通过自旋标记 EPR 进行肽-膜相互作用。

• 发表时间: 2015
• 作者: Smirnova, Tatyana I;Smirnov, Alex I
• 通讯作者: Smirnov, Alex I

Alternative Reactivity of Leucine 5-Hydroxylase Using an Olefin-Containing Substrate to Construct a Substituted Piperidine Ring.

使用含烯烃底物构建取代哌啶环的亮氨酸 5-羟化酶的替代反应性。

• 发表时间: 2020
• 影响因子: 2.9
• 作者: Cha, Lide;Milikisiyants, Sergey;Davidson, Madison;Xue, Shan;Smirnova, Tatyana;Smirnov, Ale;Guo, Yisong;Chang, Wei
• 通讯作者: Chang, Wei

Surface electrostatics of lipid bilayers by EPR of a pH-sensitive spin-labeled lipid.

通过 pH 敏感的自旋标记脂质的 EPR 分析脂质双层的表面静电。

• 发表时间: 2013-01-08
• 影响因子: 3.4
• 作者: Voinov, Maxim A;Rivera;Smirnov, Alex I
• 通讯作者: Smirnov, Alex I

A biradical-tagged phospholipid as a polarizing agent for solid-state MAS Dynamic Nuclear Polarization NMR of membrane proteins.

双自由基标记磷脂作为膜蛋白固态 MAS 动态核极化 NMR 的偏振剂。

• 发表时间: 2019
• 影响因子: 3.2
• 作者: Good, Daryl B;Voinov, Maxim A;Bolton, David;Ward, Meaghan E;Sergeyev, Ivan V;Caporini, Marc;Scheffer, Peter;Lo, Andy;Rosay, Melanie;Marek, Antonin;Brown, Leonid S;I Smirnov, Ale;Ladizhansky, Vlad
• 通讯作者: Ladizhansky, Vlad